Preservation of food products



tion and use of an antibiotic substance, and more particularly to thepreparation and use of an anbiotic substance of great potency ashereinafter l shaken at intervals. The acetone layer containa more fullydescribed. i ing the acetone-soluble constituents of the com Therefore,an object of the present invention mon agrimony was separated fromtheinsoluble is to provide a potent antibiotic substance of materialbyfiltration, and the acetone was dislant rigi tilled oiT at 42 C. Theresidue was taken up in A further object of the invention is to provideethyl alcohol (95%) so that 1 ml. of the alcoholic an antibioticsubstance which is nontoxic to extract contained the acetone-solublematerial in humans. 3 grams of the plant.

'Anadditional object of'the invention is to pro- In order to evaluatethe efiectiveness of the vide an antibiotic substance which is heatstable. antibiotic material in the alcohol solution ob- Another objectof th inventioni to employ tained in accordance with the example, thefolan antibiotic substance having an origin in a lowing test wasperformed: vegetative plant as an efiective preservative for Serialdilutions of the extract were made in 10 food du ts, cc. broth tubes andthe tubes inoculated with In achieving the objects of the presentinvenbacteria. Each 10 ml. of the broth tube contion,-thefine1ycut orground common agrimony tamed p X ma ly 75,000 cells of a foodplant-including the stems, leaves, or roots, is poisoning strain ofStaphylococcus aureus, Clostreated with a suitable organic solvent, suchas iridium spcr genes, Bac llus m'ger, Achromobacacetone. The acetonelayer containing the con-. t er perolens, and Remlcilltum. notatum. Thetubes stituents of the plant tissue soluble therein is were allowed toincubate at a temperature of 37 separated from the acetone-insolublefibrous C. and were inspected for baterial growth at the mass, theacetone .removed therefrom, and the end of 1, 2, 3, 6, and '7 days- Inthe following extract taken up in a carrier solvent, such as table aretabulated the results obtained with the ethanol. 7 aforesaid bacteria.The minus signs in said table The following example illustratespreferred indicate no bacterial growth, and the positive embodiments ofthe invention, and the tests fallsigns indicate the presence of viablebacteria at ing thereunder illustrate the potency of the matheincubation periods specified;

. e Table I Time Achro- Pen. 313: Eitii. egg; :51.; g ggggig 5bit? m;-5352 a t l 'I" Patented Apr. 24,1951

PRESERVATION OF FOOD PRODUCTS Lloyd B. J enscn, Chicago, Ill., andWilliam A.

Miller, Manhattan, Kans., assignors to Swift & Company, Chicago, 111., acorporation of Illinois No Drawing.

Application February 23, 1949, Serial No.

6 Claims. (01. 99

- The present invention relates to the preparatimicrobial substance ofplant origin.

The invention contemplates the utilization of :5 an antibiotic substancenaturally associated with tissues of the plant 'Agrimonia' gryposepala,generally= called common agrimony. We have found that the tissue of theplant yields an antiterial obtained in accordance with the processes Ofthe example.

EXAMPLE 150 grams of the stems, leaves, and roots of the common agrimonyplant were finely ground. The ground plant was placed in a flask andcovered with'acetone. The flask was held at room temperature forapproximately 24 hours and The data given in the above table indicatethat the antibiotic effects of the common agrimony plant extract vary inpotency when tested against different organisms. Thus, the extract killsStaphylococcus in dilutions as high at 1 to 2000, Bacillus niger indilutions as high as 1 to 1000, and Clostridium sporogenes in dilutionsof 1 to 100. The bactericidal properties of the extracts lethallyeffective dilutions indicated for the various organisms weredemonstrated by inoculating nutrient agar plates with 0.1 cc. of theaforesaid dilutions which had been allowed to incubate for days. After 3days incubation of the inoculated plates, no viable bacteria wereobserved as a result of the transfers.

Although acetone is the preferred primary solvent, we have found thatother organic solvents are suitable as the extracting solvent. Forexample, dioxane, ethyl acetate, methyl ethyl ke tone, and ethanol maybe used. We have also though great strides have been made in improvingthe quality and keeping time of various food products by refrigeration,canning, curing, plant sanitation, etc., the problem of preservation offood requires the constant attention of the food processor to improvelong-established methods of treating food and to devise new means offood control.

Food spoilage is due primarily to microbial action, and the species ofmicroorganism responsible for food deterioration are numerous. Thesporing genera, Clostridium and Bacillus, for example, whencontaminating food, cause food spoilage. Another serious concern of thefood handler is food poisoning, which is often of an insidious naturebecause food products may be contaminated with one or more of theharmful bacteria referred to herein and yet appear organofound that awater extraction of the antibiotic material to beonly slightly effectiveas an antibiotic because of the proteinaceous material dissolvedtherein.

The heat stability of the antibiotic material was determined by boilingportions of the said material for 3.5 minutes as indicated in Table II,and then inoculating with about 40,000 Staphylococcus cells permilliliter. Afterincubating' the material at a temperature ofapproximately 75 F. for '7 days, the, lethal potency was 1:2000.

It is obvious that many variations in the solvent treating process maybe advantageously employed. For example, since the antibiotic materialisolated from the common agrimony plant is heat stable, solventtemperatures may range from room temperature to boiling temperature ofthe solvent, with consequent variations in extraction time. The higherthe temperature, generally the less time is required for extraction.Moreover, the extraction process may be carried out in one step, forexample, with ethanol or with a mixture of solvents. We have found itpreferable to process the antibiotic substance contained in the commonagrimony plant by a two-stage solvent process involving treating thesubstance with a primary solvent which is substantially removed, andthen taking up the resulting extract with another solvent so as to forma solution, which is a convenient form of using the product. We havefound that acetone is a very effective primary solvent in producing theantibiotic substance from the plant tissue. After evaporating off theacetone, the preferred solution is made with a carrier solvent, such asethanol, which may be consumed internally without harm. By using ethylalcohol, we also utilize its leptically sound. Among the food poisoningbacteria, the most common food poisoner is Staphylococcus. I

The antibacterial substance of the present invention may be used toretard or prevent microbial growth in various food products, all ofwhich tend to harber pathogenic organisms in varying degrees. Theantibiotic material disclosed herein, therefore, may advantageously beemployed, for example, in treating meat products, such as hams,sausages, and canned meats; filledpastries, such as cream puffs,chocolate eclairs, custards, and the like; and canned vegetable toprevent swelling due to organisms, such as the genus Bacillus.

The invention will be more fully understood from the followingdescription of methods of using the antibiotic substance.

As hereinbefore mentioned, we have found that the antibiotic reagentherein described is lethal to anumber of microbes in various dilutionsof the extract, the extract being of such strength that 1 ml. of thealcoholic solution thereof contained the acetone-soluble constituents in3 grams of the plant. Therefore, the addition of 1 part of the solutionto 500 parts of the conventional pickling solution will result in a meatproduct cured therewith which has a very marked increase in resistanceto the development of pathogeni-c organisms. 1

The following is an example of a curing pickle stock which may be usedin a process for quick curing hams when provided with antibioticsubstance in the proportions outlined above. That is, the antibioticsubstance is added in an amount such as to effect a l to 500 dilution ofthe antibiotic material of the arbitrary strength herein described ('1ml.=3 g. of the plant).

To gallons of a 90 Salometer solution there are added:

'7 lbs. sodium nitrate 10 oz. sodium nitrite 20 lbs. sugar 378.5 cc.antibiotic solution meat product was placed in six 6-pound canswell-known properties, such as nontoxicity and ability to preventcontamination.

In the processing of food products, a serious problem is thepreservation thereof; and a1- and retorted to an inside temperature ofF. and incubated at 99 F. After 30 days of incubation no swelling of thecans could be observed. The control cans swelled in one week.

A further example of the utility of the antibiotic material is their usein a cream filling for various pastries. To determine the effectiveness,a standard cream filling used in eclairs was inoculated with a bacterialsolution containing 100,000 living Staphylococcus aureus S-l per 100 m1.and the filling incubated at a temperature between 75 F. and 80 F. Theresults of the tests are given in Table III.

Table III 5 Incubated at 75-80 F. Dilution Staphylococcus (8-1) perAntibiotic in cream gram filling 10 24 hrs. 48 hrs. 72 hrs.

Common Agrimony an 1-300 15,000 12,000,000 No Antibiotic 0 140,000,000No Staphylococci 0 0 0 0 The foregoing tests indicate that theantibiotic preparation exerts a pronounced inhibiting action against theStaphylococcus aureus in cream fillings.

Obviously, many modifications and variations of the invention ashereinbefore set forth may be made without departing from the spirit andscope thereof, and therefore only such limitations should be, imposed asare indicated in the appended claims.

We claim: 1. In the process of treating food products, the step ofadding to a food product a small amount of an alcoholic solution ofantibiotic substance derived from the plant Agrimoma meat, the step ofadding to said pickling solution a small amount of the antibioticsubstance derived from the plant Agrimonia gryposepala by treatment witha non-aqueous organic solvent whereby the growth of pathogenic bacteriaresponsible for food spoilage and poisoning in the meat is substantiallyretarded.

3. In the process of preparing pastries containing fillings subject tospoilage by pathogenic bacteria, the step of adding to the said fillinga small amount of the antibiotic substance derived from the plantAgrimoma gryposepala by treatment with a non-aqueous organic solventwhereby the growth of pathogenic bacteria is substantially retarded.

4. A meat pickle including a small amount of an antibiotic substancederived from the plant Agrimonia gryposepala by treatment with anonaqueous organic solvent.

5. A cream filling for pastries including a small amount of anantibiotic substance derived from the plant Agrimom'a gryposepala bytreatment with a non-aqueous organic solvent.

6. In the process of treating food products, the step of adding to afood product a small amount of an antibiotic substance derived from theplant Agrimom'a gryposepala by treatment with a non-aqueous organicsolvent to preserve said food product against pathogenic bacteriaresponsible for food poisoning and spoilage.

LLOYD B. JENSEN. WILLIAM A. MILLER.

REFERENCES CITED The following references are of record in the file ofthis patent:

UNITED STATES PATENTS Number Name Date 7 3 469,850 Chesebrough Mar. 1,1892 2,084,864 Paddock June 22, 1937 2,098,110 Schertz et a1. Nov. 2,1937 2,117,478 Hall May 17, 1938 2,180,750 Urbain Nov. 21, 1939 OTHERREFERENCES Book Publishers, Chicago, page 475.

1. IN THE PROCESS OF TREATING FOOD PRODUCTS, THE STEP OF ADDING TO AFOOD PRODUCT A SMALL AMOUNT OF AN ALCOHOLIC SOLUTON OF ANTIBIOTICSUBSTANCE DERIVED FROM THE PLANT AGRIMONIA GRYPOSEPALA TO PRESERVE SAIDFOOD PRODUCT AGAINST PATHOGENIC BACTERIA RESPONSBILE FOR FOOD POISONINGAND SPOILAGE.